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Jörg Heukelbach . Fabíola A. S. Oliveira .Richard Speare
A new shampoo based on neem (Azadirachta indica) is highlyeffective against head lice in vitro
Received: 7 February 2006 / Accepted: 10 February 2006 / Published online: 28 March 2006
Abstract Because topical compounds based on insecticid-
al chemicals are the mainstay of head lice treatment, butresistance is increasing, alternatives, such as herbs and oils
Prevalence of head lice (caused by infection with Pediculus
are being sold to treat head lice. To test a commercial
capitis) has been increasing worldwide (Burgess
shampoo based on seed extract of Azadirachta indica
Topical insecticides based on insecticidal chemicals are
(neem tree) for its in vitro effect, head lice (n=17) were
still the mainstay of head lice treatment (Heukelbach and
collected from school children in Australia and immersed
Feldmeier ), and resistance to the common over-the-
in Wash-Away Louse™ shampoo (Alpha-Biocare GmbH,
counter pediculicides has been reported repeatedly by
Germany). Vitality was evaluated for more than 3 h by
clinicians and confirmed by in vitro studies (Burkhart et al.
examination under a dissecting microscope. Positive and
negative controls were a commercially available head lice
Elston Hunter and Barker ; Mumcuoglu et al.
treatment containing permethrin 1% (n=19) and no
; Picollo et al. ; Yoon et al. ). Resistance is
treatment (n=14). All lice treated with the neem shampoo
particularly frequent in countries where head lice are
did not show any vital signs from the initial examination
common and where chemical insecticides are extensively
after immersion at 5–30 min; after 3 h, only a single louse
used, such as in the US, UK, Israel, and Australia. Because
showed minor signs of life, indicated by gut movements, a
the prevalence and degree of insecticide resistance is
mortality of 94%. In the permethrin group, mortality was
expected to increase, alternative topical therapies for head
20% at 5 min, 50% at 15 min, and 74% after 3 h. All 14
head lice of the negative control group survived during the
We tested the shampoo “Wash-Away Louse™” based on
observation period. Our data show that Wash-Away
seeds of Azadirachta indica (neem tree) for its effect on
Louse™ is highly effective in vitro against head lice. The
head lice in vitro. The results show that the product is
neem shampoo was more effective than the permethrin-
highly effective against P. capitis.
based product. We speculate that complex plant-basedcompounds will replace the well-defined chemical pedicu-licides if resistance to the commonly used products further
Head lice were collected from two school children and their
mothers in Townsville, Australia on two occasions
Department of Community Health, School of Medicine,
separated by a month. After collection, lice were pooled
and held on human hair in 5 cm diameter plastic Petri
dishes at 27°C and 50% relative humidity. In vitro tests
were started within 1 h after collection of lice, or when this
was not possible, lice were fed on the dorsum of the hand of
Tropical Medicine and Rehabilitation Sciences,
one of the investigators and used for testing within 1 h after
A commercially available head lice shampoo based on
extracts of neem seeds (Wash-Away Louse™; Alpha-
Departamento de Saúde Comunitária, Faculdade de Medicina,
Biocare GmbH, Düsseldorf, Germany) was tested.
Wash-Away Louse™ contains a patented oily extract from
seeds of the plant A. indica, formulated with the following
caprate, cocamidoprophybetaine, glycerol, lauryl gluco-
sive fashion or no righting reflex when rolled onto the back
side, dicaprylyl ether, and as preservation components
benzyl alcohol, benzoic acid, and sorbic acid.
Examinations were performed after 5, 10, 15, 20, 30, 45,
As a positive control, a 1%-permethrin-based over-the-
60, 90, and 180 min. All lice were maintained at 27°C
counter compound was used (Quellada Head Lice Treat-
For the tests, lice clasping hair strands were immersed
completely in the product for 1 min, then placed with hairs
onto Whatman filter paper in Petri dishes. To prevent licefrom desiccation, the filter paper had been previously
Using the strict criteria for mortality, all lice treated with the
moistened by 200 μl tap water. After lice were placed on
Wash-Away Louse™ shampoo did not show any vital signs
the filter paper, obvious pools of the liquid product rests
after 5 min (Fig. ). After 30 min, in two lice minimal
adherent to the body of lice were wiped from the lice by a
internal gut movements were observed, giving a mortality
jeweller’s forceps directed under a dissecting microscope.
of 88.2%. After 3 h, mortality was 94.1% with one louse
Negative control lice were placed directly on the filter
showing persistent gut movements. All 14 head lice in the
paper without any treatment. To simulate treatment on an
negative control group survived during the observation
infested host, head lice were washed in tap water after
period. In the permethrin group, the number of vital lice
20 min and placed into a new Petri dish.
decreased steadily from 20% at 5 min to 73.7% at 3 h
The number of lice was 17, 19, and 14 in the neem
(Fig. Interestingly, the activity of two lice remained
group, the permethrin group, and the control group,
normal throughout the 3-h observation period.
respectively. Lice were tested in batches of seven to 14
“Mortality” as defined by the usual criteria of loss of
righting reflex was 100% in the Wash-Away Louse™
Lice on the filter paper were examined under a dissecting
shampoo group during the observation period and ranged
microscope by a single observer in all cases to prevent
between 68 and 90% in the permethrin group (Fig. ).
The criteria used for survival of lice were extremely
strict: if any even minor signs of life, such as internal
movements, or movements of antennae or minimal legmovements were observed (with or without stimulation by
The usual criteria for “mortality” in head and body lice is
a forceps), the lice were categorized as alive. The lice were
not physical death, but a lesser state judged by the inability
judged as dead if there were no vital signs at all (complete
of the insects to walk in a progressive fashion (Mougabure
physical death). These data were compared to the usual
et al. to show a righting reflex when rolled onto its
criteria for “mortality”: the inability to walk in a progres-
back (Hunter and Barker or when movement hasceased and appendages fail to move when touched with a
Fig. 1 Mortality of head licetreated with Wash-Away Louse™
different points in time, usinghighly strict criteria for mortal-
MortalityTime (min)
different points in time, asdefined by usual criteria for
survival (inability to walk in aprogressive fashion, no righting
MortalityTime (min)
needle (Oladimeji et al. However, the use of strict
criteria (complete physical death of the insects) is
formulation has been shown to be efficacious as a repellent
particularly important, as it is known that head lice can
against head lice (Mumcuoglu et al. ). In clinical
resurrect within a short period of time (Burkhart and
studies, a combination of paw paw, thymol, and tea tree oil,
Burkhart Using usual and also strict criteria for
a combination of coconut oil, anise, and ylang ylang, and a
mortality, our data show that Wash-Away Louse™ shampoo
preparation from Annona squamosa seed extract were
is highly effective in vitro against head lice collected in
highly effective against head lice infestations (McCage et
Australia, an area where resistance to commonly used
al. ; Mumcuoglu et al. ; Tiangda et al. ).
pediculicides is prevalent. After 5 min, all head lice did not
In conclusion, it can be speculated that the tested
show any vital signs and did not recover after 3 h. The
shampoo and similar products will substitute chemical
effect of the shampoo was better than a permethrin-based
compounds on the market, if resistance to the commonly
product, which is considered the first line treatment.
used pediculicides further increases.
First data of a field study from Egypt indicate that Wash-
Away Louse™ is also highly effective for treatment of
The authors thank Chris Cahill for assistance
individuals heavily infested with head lice (Ghaffar and
in obtaining head lice. Wash-Away Louse™ neem shampoo was
provided free of charge by Alpha Biocare GmbH, Düsseldorf,
The neem tree has the widest spectrum of use of all
Germany. Jorg Heukelbach was supported by an EndeavourAustralia Research Fellowship.
natural products. The first known use of neem by theHarrappa culture in ancient India dates back 4,500 years(Dasgupta et al. ). Today, neem extracts are used to
treat various skin diseases, as an antiseptic substance,against endo- and ectoparasites, or simply as hair
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